Cancer treatment method

ABSTRACT

A method of treating cancer is described including administration of a 4-quinazolineamine and at least one other anti-neoplastic agent as well as pharmaceutical combinations and compositions containing the same.

This application is filed pursuant to 35 U.S.C. § 371 as a U.S. NationalPhase Application of International Application No. PCT/US02/01130 filedJan. 14, 2002, which claims priority from 60/262,402 filed Jan. 16,2001.

BACKGROUND OF THE INVENTION

The present invention relates to pharmaceutical combinations and methodsof treating cancer utilizing the same. Specifically, the inventionrelates to a combination of one of several quinazoline derivatives,which are inhibitors of erb-B2 and/or EGFR and other anti-neoplastics,as well as use of the combination in the treatment of cancer.

Effective treatment of hyperproliferative disorders, including cancer,is a continuing goal in the oncology field. Protein tyrosine kinasescatalyse the phosphorylation of cell growth and differentiation ofspecific tyrosyl residues in various proteins involved in the regulationof cell growth and differentiation. (A. F. Wilks, Progress in GrowthFactor Research, 1990, 2, 97–111; S. A. Courtneidge, Dev. Supp.l, 1993,57–64; J. A. Cooper, Semin. Cell Biol., 1994, 5(6), 377–387; R. F.Paulson, Semin. Immunol., 1995, 7(4), 267–277; A. C. Chan, Curr. Opin.Immunol., 1996, 8(3), 394–401). Inappropriate or uncontrolled activationof many of such kinases, i.e., aberrant protein tyrosine kinaseactivity, for example by over-expression or mutation, has been shown toresult in uncontrolled cell growth.

The erbB family of protein tyrosine kinases is one group of such kinaseswhich has been implicated in human malignancies. Elevated EGFr activityhas, for example, been implicated in non-small cell lung, bladder andhead and neck cancers, and increased c-erbB-2 activity in breast,ovarian, gastric and pancreatic cancers. Consequently, inhibition ofsuch protein tyrosine kinases should provide a treatment for disorderscharacterized by aberrant erb family protein kinase activity.

International Patent Application PCT/EP99/00048 filed Jan. 8, 1999, andpublished as WO 99/35146 on Jul. 15, 1999, discusses PTKs including erbBfamily PTKs. This published application discloses bicyclicheteroaromatic compounds, includingN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine;(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-amine;(4-(3-Fluoro-benzyloxy)-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)quinazolin-4-yl)-amineas well as hydrochloride salts thereof. These compounds show inhibitionactivity against erbB family PTKs.

Combination therapy is rapidly becoming the norm in cancer treatment,rather than the exception. Oncologists are continually looking foranti-neoplastic compounds which when utilized in combination provides amore effective and/or enhanced treatment to the individual suffering theeffects of cancer. Typically, successful combination therapy providesimproved and even synergistic effect over monotherapy.

The present inventors have now identified combinations ofchemotherapeutic agents that provide increased activity overmonotherapy. In particular, multiple drug combinations that includeinhibitors of the erbB family of kinases in combination with otheranti-neoplastic agents.

SUMMARY OF THE INVENTION

In a first aspect of the present invention, there is provided a methodof treating cancer in a mammal, including administering to said mammal atherapeutically effective amount of (a) a compound of formula I,

and salts, solvates or physiologically functional derivatives thereof,

-   wherein R₁ is Cl or Br; X is CH, N, or CF; and Het is thiazole or    furan; and-   (b) at least one anti-neoplastic agent.

In a second aspect of the present invention, there is provided apharmaceutical combination including therapeutically effective amountsof: (a) a compound of formula I and salts, solvates or physiologicallyfunctional derivatives thereof and (b) at least one anti-neoplasticagent.

In a third aspect of the present invention, there is provided apharmaceutical combination including a therapeutically effective amountof (a) a compound of formula I and salts, solvates or physiologicallyfunctional derivatives thereof and (b) at least one anti-neoplasticagent for use in therapy.

In a fourth aspect of the present invention, there is provided use of apharmaceutical combination including therapeutically effective amountsof (a) a compound of formula I and salts, solvates or physiologicallyfunctional derivatives thereof and (b) at least one anti-neoplasticagent in the preparation of a medicament for use in the treatment ofcancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts anti-tumor activity in a subcutaneous human xenograftmouse model dosed with a compound of Example 1 and carboplatinindividually and in combination versus HN5 (human head and neck tumorline).

FIG. 2 depicts anti-tumor activity in a subcutaneous human xenograftmouse model dosed with a compound of Example 1 and paclitaxel (Taxol®)individually and in combination versus BT474 (human breast tumor line).

FIG. 3 depicts anti-tumor activity in a subcutaneous human xenograftmouse model dosed with a compound of Example 1 and paclitaxelindividually and in combination versus NCl H-322(human lung tumor line).

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term “effective amount” means that amount of a drugor pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal or human that is being sought, forinstance, by a researcher or clinician. Furthermore, the term“therapeutically effective amount” means any amount which, as comparedto a corresponding subject who has not received such amount, results inimproved treatment, healing, prevention, or amelioration of a disease,disorder, or side effect, or a decrease in the rate of advancement of adisease or disorder. The term also includes within its scope amountseffective to enhance normal physiological function.

As used herein, the term “physiologically functional derivative” refersto any pharmaceutically acceptable derivative of a compound of FormulaeI, II, III, or IV, for example, an ester or an amide, which uponadministration to a mammal is capable of providing (directly orindirectly) a compound of Formulae I, II, III, or IV or an activemetabolite thereof. Such derivatives are clear to those skilled in theart, without undue experimentation, and with reference to the teachingof Burger's Medicinal Chemistry And Drug Discovery, 5^(th) Edition, Vol1: Principles and Practice, which is incorporated herein by reference tothe extent that it teaches physiologically functional derivatives.

As used herein, the term “solvate” refers to a complex of variablestoichiometry formed by a solute (in this invention, a compound offormula I, II, III, or IV or a salt or physiologically functionalderivative thereof) and a solvent. Such solvents for the purpose of theinvention may not interfere with the biological activity of the solute.Examples of suitable solvents include, but are not limited to, water,methanol, ethanol and acetic acid. Preferably the solvent used is apharmaceutically acceptable solvent. Examples of suitablepharmaceutically acceptable solvents include water, ethanol and aceticacid. Most preferably the solvent used is water.

The compounds of formulae I, II, III and IV have the ability tocrystallize in more than one form, a characteristic, which is known aspolymorphism, and it is understood that such polymorphic forms(“polymorphs”) are within the scope of formulae I, II, III and IV.Polymorphism generally can occur as a response to changes in temperatureor pressure or both and can also result from variations in thecrystallization process. Polymorphs can be distinguished by variousphysical characteristics known in the art such as x-ray diffractionpatterns, solubility, and melting point.

Typically, the salts of the compounds of formula I, II, III, or IV arepharmaceutically acceptable salts. Salts encompassed within the term“pharmaceutically acceptable salts” refer to non-toxic salts of thecompounds of this invention. Salts of the compounds of formula I, II,III, or IV may comprise acid addition salts derived from a nitrogen on asubstituent in the compound of formula I, II, III, or IV. Representativesalts include the following salts: acetate, benzenesulfonate, benzoate,bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate,camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride,edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate,glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate,lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, monopotassium maleate,mucate, napsylate, nitrate, N-methylglucamine, oxalate, pamoate(embonate), palmitate, pantothenate, phosphate/diphosphate,polygalacturonate, potassium, salicylate, sodium, stearate, subacetate,succinate, tannate, tartrate, teoclate, tosylate and ditosylate,triethiodide, trimethylammonium and valerate. Other salts, which are notpharmaceutically acceptable, may be useful in the preparation ofcompounds of this invention and these form a further aspect of theinvention. Furthermore, such salt may be in anhydrous or hydrated form.In one embodiment, the compound of formula I, II, III, or IV is ahydrochloride or ditosylate salt, preferably a ditosylate salt, morepreferably the monohydrate of the ditosylate salt.

The side chain CH₃SO₂CH₂CH₂NHCH₂ of the compounds of formula I, II, III,or IV may be linked to any suitable position of the group Het Similarly,the phenyl group of the quinazoline core may be linked to any suitableposition of the group Het.

As recited above, a method of treating cancer is provided which includesadministering therapeutically effective amounts of a compound of formulaI and salts, solvates or physiologically functional derivatives thereofand at least one anti-neoplastic agent. The compound of formula I andsalts, solvates or physiologically functional derivatives thereof are asdefined above, that is R₁ is Cl or Br; X is CH, N, or CF; and Het isthiazole or furan.

It is to be understood that reference to compounds of formulae I above,and II, III, and IV following herein refers to compounds within thescope of these formulae as defined above unless specifically limited insubsequent reference to such formula with respect to R₁, Het, and X. Itis also understood that the embodiments of the present inventiondescribed herein, including uses and compositions, are applicable to notonly formula I, but to formula II, III, or IV, unless specificallystated otherwise.

In one embodiment, R₁ is Cl; X is CH; and Het is furan, preferably acompound of Formula II and salts, solvates or physiologically functionalderivatives thereof.

The compound of formula II has the chemical nameN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine.

In another embodiment, R₁ is Cl; X is CH; and Het is thiazole,preferably a compound of formula III and salts, solvates orphysiologically functional derivatives thereof.

The compound of formula III is(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-amine.

In a further embodiment, R₁ is Br; X is CH; and Het is furan,preferably, a compound of formula IV and salts, solvates orphysiologically functional derivatives thereof.

The compound of formula IV is(4(3-Fluoro-benzyloxy)-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)quinazolin-4-yl)-amine.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be utilized in the cancer treatmentmethod of the present invention. Typical anti-neoplastic agents usefulin the present invention include, but are not limited to,anti-microtubule agents such as diterpenoids and vinca alkaloids;platinum coordination complexes; alkylating agents such as nitrogenmustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, andtriazenes; antibiotic agents such as anthracyclins, actinomycins andbleomycins; topoisomerase II inhibitors such as epipodophyllotoxins;antimetabolites such as purine and pyrimidine analogues and anti-folatecompounds; topoisomerase I inhibitors such as camptothecins; hormonesand hormonal analogues; signal transduction pathway inhibitors;non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeuticagents; proapoptotic agents; and cell cycle signaling inhibitors.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7,β10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem, Soc., 93:2325. 1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA,77:1561–1565 (1980); Schiff et al., Nature, 277:665–667 (1979); Kumar,J. Biol, Chem, 256: 10435–10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LeQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219–235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaleJournal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann.Intem, Med., 111:273, 1989) and for the treatment of breast cancer(Holmes et al., J. Nat. Cancer Inst., 83:1797, 1991.) It is a potentialcandidate for treatment of neoplasms in the skin (Einzig et. al., Proc.Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastireet. al., Sem. Oncol., 20:56, 1990). The compound also shows potentialfor the treatment of polycystic kidney disease (Woo et. al., Nature,368:750. 1994), lung cancer and malaria. Treatment of patients withpaclitaxel results in bone marrow suppression (multiple cell lineages,Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide, 1998) related tothe duration of dosing above a threshold concentration (50 nM) (Kearns,C. M. et. al., Seminars in Oncology, 3(6) p. 16–23, 1995).

Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine,N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkle plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine,3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate(1:2)(salt)],commercially available as an injectable solution of vinorelbine tartrate(NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine isindicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo, aquation and form intra- and interstrandcrosslinks with DNA causing adverse biological effects to the tumor.Examples of platinum coordination complexes include, but are not limitedto, cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′],is commercially available as PARAPLATIN® as an injectable solution.Carboplatin is primarily indicated in the first and second linetreatment of advanced ovarian carcinoma. Bone marrow suppression is thedose limiting toxicity of carboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-resectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also know as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins. Epipodophyllotoxins are phase specificanti-neoplastic agents derived from the mandrake plant.Epipodophyllotoxins typically affect cells in the S and G₂ phases of thecell cycle by forming a ternary complex with topoisomerase II and DNAcausing DNA strand breaks. The strand breaks accumulate and cell deathfollows. Examples of epipodophyllotoxins include, but are not limitedto, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leucopenia tends to be more severethan thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leucopenia and thrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.

5-fluorouracil,5-fluoro-2,4-(1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2 (1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leucopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of thioguanine administration.However, gastrointestinal side effects occur and can be dose limiting.Other purine analogs include pentostatin, erythrohydroxynonyladenine,fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of gemcitabine administration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leucopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl,(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®. Irinotecan is a derivative of camptothecin which binds,along with its active metabolite SN-38, to the topoisomerase I—DNAcomplex. It is believed that cytotoxicity occurs as a result ofirreparable double strand breaks caused by interaction of thetopoisomerase I:DNA: irintecan or SN-38 ternary complex with replicationenzymes. Irinotecan is indicated for treatment of metastatic cancer ofthe colon or rectum. The dose limiting side effects of irinotecan HClare myelosuppression, including neutropenia, and GI effects, includingdiarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dionemonohydrochloride, is commercially available as the injectable solutionHYCAMTIN®. Topotecan is a derivative of camptothecin which binds to thetopoisomerase I—DNA complex and prevents religation of singles strandbreaks caused by Topoisomerase I in response to torsional strain of theDNA molecule. Topotecan is indicated for second line treatment ofmetastatic carcinoma of the ovary and small cell lung cancer. The doselimiting side effect of topotecan HCl is myelosuppression, primarilyneutropenia.

Also of interest, is the camptothecin derivative of formula A following,currently under development, including the racemic mixture (R,S) form aswell as the R and S enantiomers:

known by the chemical name“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R,S)-camptothecin(racemic mixture) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R)-camptothecin(R enantiomer) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(S)-camptothecin(S enantiomer). Such compound as well as related compounds aredescribed, including methods of making, in U.S. Pat. Nos. 6,063,923;5,342,947; 5,559,235; 5,491,237 and pending U.S. patent application Ser.No. 08/977,217 filed Nov. 24, 1997.

Hormones and hormonal analogues are useful compounds for treatingcancers in which there is a relationship between the hormone(s) andgrowth and/or lack of growth of the cancer. Examples of hormones andhormonal analogues useful in cancer treatment include, but are notlimited to, adrenocorticosteroids such as prednisone and prednisolonewhich are useful in the treatment of malignant lymphoma and acuteleukemia in children; aminoglutethimide and other aromatase inhibitorssuch as anastrozole, letrazole, vorazole, and exemestane useful in thetreatment of adrenocortical carcinoma and hormone dependent breastcarcinoma containing estrogen receptors; progestrins such as megestrolacetate useful in the treatment of hormone dependent breast cancer andendometrial carcinoma; estrogens, androgens, and anti-androgens such asflutamide, nilutamide, bicalutamide, cyproterone acetate and5α-reductases such as finasteride and dutasteride, useful in thetreatment of prostatic carcinoma and benign prostatic hypertrophy;anti-estrogens such as tamoxifen, toremifene, raloxifene, droloxifene,iodoxyfene, as well as selective estrogen receptor modulators (SERMS)such those described in U.S. Pat. Nos. 5,681,835, 5,877,219, and6,207,716, useful in the treatment of hormone dependent breast carcinomaand other susceptible cancers; and gonadotropin-releasing hormone (GnRH)and analogues thereof which stimulate the release of leutinizing hormone(LH) and/or follicle stimulating hormone (FSH) for the treatmentprostatic carcinoma, for instance, LHRH agonists and antagagonists suchas goserelin acetate and luprolide.

Signal transduction pathway inhibitors are those inhibitors, which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation. Signaltranduction inhibitors useful in the present invention includeinhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases,SH2/SH3domain blockers, serine/threonine kinases, phosphotidylinositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

Several protein tyrosine kinases catalyse the phosphorylation ofspecific tyrosyl residues in various proteins involved in the regulationof cell growth. Such protein tyrosine kinases can be broadly classifiedas receptor or non-receptor kinases.

Receptor tyrosine kinases are transmembrane proteins having anextracellular ligand binding domain, a transmembrane domain, and atyrosine kinase domain. Receptor tyrosine kinases are involved in theregulation of cell growth and are generally termed growth factorreceptors. Inappropriate or uncontrolled activation of many of thesekinases, i.e. aberrant kinase growth factor receptor activity, forexample by over-expression or mutation, has been shown to result inuncontrolled cell growth. Accordingly, the aberrant activity of suchkinases has been linked to malignant tissue growth. Consequently,inhibitors of such kinases could provide cancer treatment methods.Growth factor receptors include, for example, epidermal growth factorreceptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2,erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosinekinase with immunoglobulin-like and epidermal growth factor homologydomains (TIE-2), insulin growth factor—I (IGFI) receptor, macrophagecolony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growthfactor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin(eph) receptors, and the RET protooncogene. Several inhibitors of growthreceptors are under development and include ligand antagonists,antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.Growth factor receptors and agents that inhibit growth factor receptorfunction are described, for instance, in Kath, John C., Exp. Opin. Ther.Patents (2000) 10(6):803–818; Shawver et al DDT Vol 2, No. 2 February1997; and Lofts, F. J. et al, “Growth factor receptors as targets”, NewMolecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr,David, CRC press 1994, London.

Tyrosine kinases, which are not growth factor receptor kinases aretermed non-receptor tyrosine kinases. Non-receptor tyrosine kinasesuseful in the present invention, which are targets or potential targetsof anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focaladhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Suchnon-receptor kinases and agents which inhibit non-receptor tyrosinekinase function are described in Sinh, S. and Corey, S. J., (1999)Journal of Hematotherapy and Stem Cell Research 8 (5): 465–80; andBolen, J. B., Brugge, J. S., (1997) Annual review of Immunology. 15:371–404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3domain bindingin a variety of enzymes or adaptor proteins including, PI3-K p85subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2) andRas-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussedin Smithgall, T. E. (1995), Journal of Pharmacological and ToxicologicalMethods. 34(3) 125–32.

Inhibitors of Serine/Threonine Kinases including MAP kinase cascadeblockers which include blockers of Raf kinases (rafk), Mitogen orExtracellular Regulated Kinase (MEKs), and Extracellular RegulatedKinases (ERKs); and Protein kinase C family member blockers includingblockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).IkB kinase family (IKKa, IKKb), PKB family kinases, akt kinase familymembers, and TGF beta receptor kinases. Such Serine/Threonine kinasesand inhibitors thereof are described in Yamamoto, T., Taya, S.,Kaibuchi, K., (1999), Journal of Biochemistry. 126 (5) 799–803; Brodt,P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60.1101–1107; Massague, J., Weis-Garcia, F. (1996) Cancer Surveys.27:41–64; Philip, P. A., and Harris, A. L. (1995), Cancer Treatment andResearch. 78: 3–27, Lackey, K. et al Bioorganic and Medicinal ChemistryLetters, (10), 2000, 223–226; U.S. Pat. No. 6,268,391; andMartinez-Iacaci, L., et al, Int. J. Cancer (2000), 88(1), 44–52.

Inhibitors of Phosphotidyl inositol-3 Kinase family members includingblockers of PI3-kinase, ATM, DNA-PK, and Ku are also useful in thepresent invention. Such kinases are discussed in Abraham, R. T. (1996),Current Opinion in Immunology. 8 (3) 412–8; Canman, C. E., Lim, D. S.(1998), Oncogene 17 (25) 3301–3308; Jackson, S. P. (1997), InternationalJournal of Biochemistry and Cell Biology. 29 (7):935–8; and Zhong, H. etal, Cancer res, (2000) 60(6), 1541–1545.

Also useful in the present invention are Myo-inositol signalinginhibitors such as phospholipase C blockers and Myoinositol analogues.Such signal inhibitors are described in Powis, G., and Kozikowski A.,(1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workmanand David Kerr, CRC press 1994, London.

Another group of signal transduction pathway inhibitors are inhibitorsof Ras Oncogene. Such inhibitors include inhibitors offarnesyltransferase, geranyl-geranyl transferase, and CAAX proteases aswell as anti-sense oligonucleotides, ribozymes and immunotherapy. Suchinhibitors have been shown to block ras activation in cells containingwild type mutant ras, thereby acting as antiproliferation agents. Rasoncogene inhibition is discussed in Scharovsky, O. G., Rozados, V. R.,Gervasoni, S. I. Matar, P. (2000), Journal of Biomedical Science. 7(4)292–8; Ashby, M. N. (1998), Current Opinion in Lipidology. 9 (2) 99–102;and BioChim. Biophys. Acta, (19899) 1423(3):19–30.

As mentioned above, antibody antagonists to receptor kinase ligandbinding may also serve as signal transduction inhibitors. This group ofsignal transduction pathway inhibitors includes the use of humanizedantibodies to the extracellular ligand binding domain of receptortyrosine kinases. For example Imclone C225 EGFR specific antibody (seeGreen, M. C. et al, Monoclonal Antibody Therapy for Solid Tumors, CancerTreat. Rev., (2000), 26(4), 269–286); Herceptin® erbB2 antibody (seeTyrosine Kinase Signalling in Breast cancer:erbB Family ReceptorTyrosine Kniases, Breast cancer Res., 2000, 2(3), 176–183); and 2CBVEGFR2 specific antibody (see Brekken, R. A. et al, Selective Inhibitionof VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumorgrowth in mice, Cancer Res. (2000) 60, 5117–5124).

Non-receptor kinase angiogenesis inhibitors may also find use in thepresent invention. Inhibitors of angiogenesis related VEGFR and TIE2 arediscussed above in regard to signal transduction inhibitors (bothreceptors are receptor tyrosine kinases). Angiogenesis in general islinked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR havebeen shown to inhibit angiogenesis, primarily VEGF expression. Thus, thecombination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesismakes sense. Accordingly, non-receptor tyrosine kinase inhibitors may beused in combination with the EGFR/erbB2 inhibitors of the presentinvention. For example, anti-VEGF antibodies, which do not recognizeVEGFR (the receptor tyrosine kinase), but bind to the ligand; smallmolecule inhibitors of integrin (alpha_(v) beta₃) that will inhibitangiogenesis; endostatin and angiostatin (non-RTK) may also prove usefulin combination with the disclosed erb family inhibitors. (See Bruns C Jet al (2000), Cancer Res., 60: 2926–2935; Schreiber A B, Winkler M E,and Derynck R. (1986), Science, 232: 1250–1253; Yen L et al. (2000),Oncogene 19: 3460–3469).

Agents used in immunotherapeutic regimens may also be useful incombination with the compounds of formula (I). There are a number ofimmunologic strategies to generate an immune response against erbB2 orEGFR. These strategies are generally in the realm of tumor vaccinations.The efficacy of immunologic approaches may be greatly enhanced throughcombined inhibition of erbB2/EGFR signaling pathways using a smallmolecule inhibitor. Discussion of the immunologic/tumor vaccine approachagainst erbB2/EGFR are found in Reilly R T et al. (2000), Cancer Res.60: 3569–3576; and Chen Y, Hu D, Eling D J, Robbins J, and Kipps T J.(1998), Cancer Res. 58: 1965–1971.

Agents used in proapoptotic regimens (e.g., bcl-2 antisenseoligonucleotides) may also be used in the combination of the presentinvention. Members of the Bcl-2 family of proteins block apoptosis.Upregulation of bcl-2 has therefore been linked to chemoresistance.Studies have shown that the epidermal growth factor (EGF) stimulatesanti-apoptotic members of the bcl-2 family (i.e., mcl-1). Therefore,strategies designed to downregulate the expression of bcl-2 in tumorshave demonstrated clinical benefit and are now in Phase II/III trials,namely Genta's G3139 bcl-2 antisense oligonucleotide. Such proapoptoticstrategies using the antisense oligonucleotide strategy for bcl-2 arediscussed in Water J S et al. (2000), J. Clin. Oncol. 18: 1812–1823; andKitada S et al. (1994), Antisense Res. Dev. 4: 71–79.

Cell cycle signalling inhibitors inhibit molecules involved in thecontrol of the cell cycle. A family of protein kinases called cyclindependent kinases (CDKs) and their interaction with a family of proteinstermed cyclins controls progression through the eukaryotic cell cycle.The coordinate activation and inactivation of different cyclin/CDKcomplexes is necessary for normal progression through the cell cycle.Several inhibitors of cell cycle signalling are under development. Forinstance, examples of cyclin dependent kinases, including CDK2, CDK4,and CDK6 and inhibitors for the same are described in, for instance,Rosania et al, Exp. Opin. Ther. Patents (2000) 10(2):215–230.

In one embodiment, the cancer treatment method of the claimed inventionincludes a compound of formula I and salts, solvates or physiologicallyfunctional derivatives thereof and at least one anti-neoplastic agentselected from the group consisting essentially of anti-microtubuleagents, platinum coordination complexes, alkylating agents, antibioticagents, topoisomerase II inhibitors, antimetabolites, topoisomerase Iinhibitors, hormones and hormonal analogues, signal transduction pathwayinhibitors, non-receptor tyrosine kinase angiogenesis inhibitors,immunotherapeutic agents, proapoptotic agents, and cell cycle signalinginhibitors.

In another embodiment, the cancer treatment method of the claimedinvention includes a compound of formula I and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent which is an anti-microtubule agent selected fromditerpenoids and vinca alkaloids.

In a preferred embodiment, the cancer treatment method of the claimedinvention includes a compound of formula I and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a diterpenoid.

In an alternative preferred embodiment, the cancer treatment method ofthe claimed invention includes a compound of formula I and salts,solvates or physiologically functional derivatives thereof and at leastone anti-neoplastic agent, which is a vinca alkaloid.

In another preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula I and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a platinum coordination complex.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula I and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent which is selected from the group consisting ofpaclitaxel, carboplatin, or vinorelbine.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula I and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is carboplatin.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula I and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is vinorelbine.

In a most preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula I and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is paclitaxel.

In one embodiment, the cancer treatment method of the claimed inventionincludes a compound of formula II and salts, solvates or physiologicallyfunctional derivatives thereof and at least one anti-neoplastic agentselected from the group consisting essentially of anti-microtubuleagents, platinum coordination complexes, alkylating agents, antibioticagents, topoisomerase II inhibitors, antimetabolites, topoisomerase Iinhibitors, hormones and hormonal analogues, signal transduction pathwayinhibitors, non-receptor tyrosine kinase angiogenesis inhibitors,immunotherapeutic agents, proapoptotic agents, and cell cycle signalinginhibitors.

In another embodiment, the cancer treatment method of the claimedinvention includes a compound of formula II and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent which is an anti-microtubule agent selected fromditerpenoids and vinca alkaloids.

In a preferred embodiment, the cancer treatment method of the claimedinvention includes a compound of formula II and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a diterpenoid.

In an alternative preferred embodiment, the cancer treatment method ofthe claimed invention includes a compound of formula II and salts,solvates or physiologically functional derivatives thereof and at leastone anti-neoplastic agent, which is a vinca alkaloid.

In another preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula II and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a platinum coordination complex.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula II and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent which is selected from the group consisting ofpaclitaxel, carboplatin, or vinorelbine.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula II and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is carboplatin.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula II and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is vinorelbine.

In a most preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula II and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is paclitaxel.

In one embodiment, the cancer treatment method of the claimed inventionincludes a compound of formula III and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent selected from the group consisting essentially ofanti-microtubule agents, platinum coordination complexes, alkylatingagents, antibiotic agents, topoisomerase II inhibitors, antimetabolites,topoisomerase I inhibitors, hormones and hormonal analogues, signaltransduction pathway inhibitors, non-receptor tyrosine kinaseangiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents,and cell cycle signaling inhibitors.

In another embodiment, the cancer treatment method of the claimedinvention includes a compound of formula III and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent which is an anti-microtubule agent selected fromditerpenoids and vinca alkaloids.

In a preferred embodiment, the cancer treatment method of the claimedinvention includes a compound of formula III and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a diterpenoid.

In an alternative preferred embodiment, the cancer treatment method ofthe claimed invention includes a compound of formula III and salts,solvates or physiologically functional derivatives thereof and at leastone anti-neoplastic agent, which is a vinca alkaloid.

In another preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula III and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a platinum coordination complex.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula III and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent which is selected from the group consisting ofpaclitaxel, carboplatin, or vinorelbine.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula III and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is carboplatin.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula III and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is vinorelbine.

In a most preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula III and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is paclitaxel.

In one embodiment, the cancer treatment method of the claimed inventionincludes a compound of formula IV and salts, solvates or physiologicallyfunctional derivatives thereof and at least one anti-neoplastic agentselected from the group consisting essentially of anti-microtubuleagents, platinum coordination complexes, alkylating agents, antibioticagents, topoisomerase II inhibitors, antimetabolites, topoisomerase Iinhibitors, hormones and hormonal analogues, signal transduction pathwayinhibitors, non-receptor tyrosine kinase angiogenesis inhibitors,immunotherapeutic agents, proapoptotic agents, and cell cycle signalinginhibitors.

In another embodiment, the cancer treatment method of the claimedinvention includes a compound of formula IV and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent which is an anti-microtubule agent selected fromditerpenoids and vinca alkaloids.

In a preferred embodiment, the cancer treatment method of the claimedinvention includes a compound of formula IV and salts, solvates orphysiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a diterpenoid.

In an alternative preferred embodiment, the cancer treatment method ofthe claimed invention includes a compound of formula IV and salts,solvates or physiologically functional derivatives thereof and at leastone anti-neoplastic agent, which is a vinca alkaloid.

In another preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula IV and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is a platinum coordination complex.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula IV and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent which is selected from the group consisting ofpaclitaxel, carboplatin, or vinorelbine.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula IV and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is carboplatin.

In a more preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula IV and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is vinorelbine.

In a most preferred embodiment, the cancer treatment method of theclaimed invention includes a compound of formula IV and salts, solvatesor physiologically functional derivatives thereof and at least oneanti-neoplastic agent, which is paclitaxel.

The compounds of the Formula I, including compounds of formulae II, III,and IV, or salts, solvates, or physiologically functional derivativesthereof and the at least one anti-neoplastic agent may be employed incombination concomitantly or sequentially in any therapeuticallyappropriate combination. The combination may be employed in combinationin accordance with the invention by administration concomitantly in (1)a unitary pharmaceutical composition including both compounds or (2)separate pharmaceutical compositions each including one of thecompounds. Alternatively, the combination may be administered separatelyin a sequential manner wherein one is administered first and the othersecond or vice versa. Such sequential administration may be close intime or remote in time.

The cancer treatment method of the present invention may also includeadministration of at least one additional cancer treatment therapy incombination concomitantly or sequentially in any therapeuticallyappropriate combination with the combinations of the present invention.The additional cancer treatment therapy may include radiation therapy,surgical therapy and/or at least one additional chemotherapeutic therapyincluding administration of at least one additional anti-neoplasticagent.

As recited above, a pharmaceutical combination including compounds ofthe Formula I, including compounds of formulae II, III, IV, or salts,solvates, or physiologically functional derivatives thereof and the atleast one anti-neoplastic agent is provided for in the presentinvention. Such compounds of formulae I, II, III, and IV and the atleast one anti-neoplastic agent are as described above and may beutilized in any of the combinations described above in the method oftreating cancer of the present invention.

While it is possible that, for use in therapy, therapeutically effectiveamounts of compounds of formula I, II, III, IV as well as salts,solvates and physiologically function derivatives thereof and the atleast one anti-neoplastic agent, may be administered as the rawchemical, it is possible to present the active ingredient as apharmaceutical composition. As indicated above, such elements of thepharmaceutical combination utilized may be presented in separatepharmaceutical compositions or formulated together in one pharmaceuticalformulation. Accordingly, the invention further provides a combinationof pharmaceutical compositions one of which includes therapeuticallyeffective amounts of compounds of the formula I and salts, solvates, andphysiologically functional derivatives thereof, and one or morepharmaceutically acceptable carriers, diluents, or excipients and apharmaceutical composition containing at least one anti-neoplastic agentand one or more pharmaceutically acceptable carriers, diluents, orexcipients.

Alternatively, a pharmaceutical composition is provided which includestherapeutically effective amounts of a compound of the formula I andsalts, solvates, and physiologically functional derivatives thereof, atleast one anti-neoplastic agent and one or more pharmaceuticallyacceptable carriers, diluents, or excipients. The compounds of theformula I, including compounds of formulae II, III, and IV, and salts,solvates, and physiologically functional derivatives thereof, and the atleast one anti-neoplastic agent are as described above and may beutilized in any of the combinations described above in the method oftreating cancer of the present invention.

The carrier(s), diluent(s) or excipient(s) must be acceptable in thesense of being compatible with the other ingredients of the formulationand not deleterious to the recipient thereof. According to anotheraspect of the invention there is also provided a process for thepreparation of a pharmaceutical formulation including admixing acompound of the formula I, or salts, solvates, or physiologicallyfunctional derivatives thereof, and/or at least one anti-neoplasticagent with one or more pharmaceutically acceptable carriers, diluents orexcipients.

The components of the pharmaceutical compositions, of the presentinvention, may be formulated for administration by any route, and theappropriate route will depend on the specific cancer being treated aswell as the subjects to be treated. Suitable pharmaceutical formulationsinclude those for oral, rectal, nasal, topical (including buccal,sub-lingual, and transdermal), vaginal or parenteral (includingintramuscular, sub-cutaneous, intravenous, and directly into theaffected tissue) administration or in a form suitable for administrationby inhalation or insufflation. The formulations may, where appropriate,be conveniently presented in discrete dosage units and may be preparedby any of the methods well know in the pharmacy art.

Pharmaceutical formulations adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilliquid emulsions.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water and the like. Powders are prepared by comminuting thecompound to a suitable fine size and mixing with a similarly comminutedpharmaceutical carrier such as an edible carbohydrate, as, for example,starch or mannitol. Flavoring, preservative, dispersing and coloringagents can also be present.

Capsules are made by preparing a powder mixture as described above, andfilling formed gelatin sheaths. Glidants and lubricants such ascolloidal silica, talc, magnesium stearate, calcium stearate or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugarssuch as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth or sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride and the like. Disintegrators include, without limitation,starch, methylcellulose, agar, bentonite, xanthan gum and the like.Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant andpressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, analiginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt and/oran absorption agent such as bentonite, kaolin or dicalcium phosphate.The powder mixture can be granulated by wetting with a binder such assyrup, starch paste, acadia mucilage or solutions of cellulosic orpolymeric materials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through the tablet machineand the result is imperfectly formed slugs broken into granules. Thegranules can be lubricated to prevent sticking to the tablet formingdies by means of the addition of stearic acid, a stearate salt, talc ormineral oil. The lubricated mixture is then compressed into tablets. Thecompounds of the present invention can also be combined with a freeflowing inert carrier and compressed into tablets directly without goingthrough the granulating or slugging steps. A clear or opaque protectivecoating consisting of a sealing coat of shellac, a coating of sugar orpolymeric material and a polish coating of wax can be provided.Dyestuffs can be added to these coatings to distinguish different unitdosages.

Oral fluids such as solution, syrups and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Syrups can be prepared by dissolving thecompound in a suitably flavored aqueous solution, while elixirs areprepared through the use of a non-toxic alcoholic vehicle. Suspensionscan be formulated by dispersing the compound in a non-toxic vehicle.Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols andpolyoxy ethylene sorbitol ethers, preservatives, flavor additive such aspeppermint oil or natural sweeteners or saccharin or other artificialsweeteners, and the like can also be added.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

The components of the pharmaceutical compositions of the presentinvention can also be administered in the form of liposome deliverysystems, such as small unilamellar vesicles, large unilamellar vesiclesand multilamellar vesicles. Liposomes can be formed from a variety ofphospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

The components of the pharmaceutical compositions of the presentinvention may also be delivered by the use of monoclonal antibodies asindividual carriers to which the compound molecules are coupled. Thecompounds may also be coupled with soluble polymers as targetable drugcarriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysinesubstituted with palmitoyl residues. Furthermore, the compounds may becoupled to a class of biodegradable polymers useful in achievingcontrolled release of a drug, for example, polylactic acid, polepsiloncaprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,polydihydropyrans, polycyanoacrylates and cross-linked or amphipathicblock copolymers of hydrogels.

Pharmaceutical formulations adapted for transdermal administration maybe presented as discrete patches intended to remain in intimate contactwith the epidermis of the recipient for a prolonged period of time. Forexample, the active ingredient may be delivered from the patch byiontophoresis as generally described in Pharmaceutical Research, 3(6),318 (1986).

Pharmaceutical formulations adapted for topical administration may beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols or oils.

For treatments of the eye or other external tissues, for example mouthand skin, the formulations are preferably applied as a topical ointmentor cream. When formulated in an ointment, the active ingredient may beemployed with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredient may be formulated in a cream withan oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations adapted for topical administrations to theeye include eye drops wherein the active ingredient is dissolved orsuspended in a suitable carrier, especially an aqueous solvent.

Pharmaceutical formulations adapted for topical administration in themouth include lozenges, pastilles and mouthwashes.

Pharmaceutical formulations adapted for rectal administration may bepresented as suppositories or as enemas.

Pharmaceutical formulations adapted for nasal administration wherein thecarrier is a solid include a coarse powder having a particle size forexample in the range 20 to 500 microns which is administered in themanner in which snuff is taken, i.e. by rapid inhalation through thenasal passage from a container of the powder held close up to the nose.Suitable formulations wherein the carrier is a liquid, foradministration as a nasal spray or as nasal drops, include aqueous oroil solutions of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalationinclude fine particle dusts or mists, which may be generated by means ofvarious types of metered, dose pressurised aerosols, nebulizers orinsufflators.

Pharmaceutical formulations adapted for vaginal administration may bepresented as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe formulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations may include other agents conventionalin the art having regard to the type of formulation in question, forexample those suitable for oral administration may include flavouringagents.

A therapeutically effective amount of the components of thepharmaceutical compositions of the present invention will depend on anumber of factors including, but not limited to, the age and weight ofthe mammal, the precise disorder requiring treatment and its severity,the nature of the formulation, and the route of administration, and willultimately be at the discretion of the attendant physcian orveternarian. Typically, the components of the pharmaceuticalcompositions of the present invention will be given for treatment in therange of 0.1 to 100 mg/kg body weight of recipient (mammal) per day andmore usually in the range of 1 to 10 mg/kg body weight per day.Acceptable daily dosages, may be from about 0.1 to about 1000 mg/day,and preferably from about 0.1 to about 100 mg/day.

The pharmaceutical combinations and compositions, including compounds offormula I and salts, solvates, and physiologically functionalderivatives thereof and at least one anti-neoplastic agent, describedabove, are useful in therapy and in the preparation of medicaments fortreating cancer in a mammal.

In one embodiment, the mammal in the methods and uses of the presentinvention is a human.

The following examples are intended for illustration only and are notintended to limit the scope of the invention in any way. The physicaldata given for the compounds exemplified is consistent with the assignedstructure of those compounds.

EXAMPLES

¹H NMR spectra were obtained at 500 MHz on a Bruker AMX500spectrophotometer, on a Bruker spectrophotometer at 300 MHz, on a BrukerAC250 or Bruker AM250 spectrophotometer at 250 MHz and on a Varian UnityPlus NMR spectrophotometer at 300 or 400 MHz. J values are given in Hz.Mass spectra were obtained on one of the following machines: VGMicromass Platform (electrospray positive or negative), HP5989A Engine(thermospray positive) or Finnigan-MAT LCQ (ion trap) mass spectrometer.Analytical thin layer chromatography (tic) was used to verify the purityof some intermediates which could not be isolated or which were toounstable for full characterization, and to follow the progress ofreactions. Unless otherwise stated, this was done using silica gel(Merck Silica Gel 60 F254).

The free base, HCl salts, and ditosylate salts of the compounds offormulae (I), (II), (III), and (IV), may be prepared according to theprocedures of the International Patent Application No. PCT/EP99/00048,filed Jan. 8, 1999, and published as WO 99/35146 on Jul. 15, 1999 and ofco-pending U.S. Provisional application No. 60/215,508 filed Jun. 30,2000. Such applications are incorporated herein by reference to theextent they teach the preparation of the compounds of formula (I), (II),(III), and (IV) and salts thereof. Some of such procedures are recitedagain herein as well as additional variations and procedures.

General Procedures

(A) Reaction of an Amine with a Bicyclic Species Containing a4-chloropyrimidine or 4-chloropyridine Ring

The optionally substituted bicyclic species and the specified amine weremixed in an appropriate solvent (typically acetonitrile unless otherwisespecified, although ethanol, 2-propanol or DMSO may also be used), andheated to reflux. When the reaction was complete (as judged by tlc), thereaction mixture was allowed to cool. The resulting suspension wasdiluted, e.g. with acetone, and the solid collected by filtration,washing e.g. with excess acetone, and dried at 60° C. in vacuo, givingthe product as the hydrochloride salt. If the free base was required(e.g. for further reaction), this was obtained by treatment with a basee.g. triethylamine; purification by chromatography was then performed ifrequired.

(B) Reaction of a Product from Procedure (A) with a Heteroaryl TinReagent

A stirred mixture of the product from (A), (containing a suitableleaving group such as chloro, bromo, iodo or triflate), a heteroarylstannane and a suitable palladium catalyst, such asbis(triphenylphosphine)palladium (II) chloride or1,4-bis(diphenylphosphino)butane palladium (II) chloride (prepared asdescribed in C. E. Housecroft et. al., Inorg. Chem., (1991), 30(1),125–130), together with other appropriate additives (such asdiisopropylethylamine or lithium chloride), were heated at reflux in drydioxane or another suitable solvent (e.g. DMF) under nitrogen until thereaction was complete. The resulting mixture was generally purified bychromatography on silica.

(C) Removal of a 1,3-dioxolan-2-yl Protecting Group to Liberate anAldehyde

The compound containing the 1,3-dioxolan-2-yl group was suspended in anappropriate solvent, e.g., THF and treated with hydrochloric acid,either as an aqueous solution (e.g. 2N) or as a solution in dioxane(e.g. 4 molar) and stirred at ambient temperature until the reaction wasjudged complete (e.g. by tlc or LC/MS analysis). Generally the mixturewas diluted with water, and the resulting precipitate was collected byfiltration, washed with water and dried to give the aldehyde.

(D) Reaction of an Aldehyde with an Amine by Reductive Amination

An aldehyde (such as the product of C) and the required primary orsecondary amine were stirred together in a suitable solvent (such asdichloromethane) containing glacial acetic acid (4A molecular sieves mayalso be present) for ca. 1 h. A suitable reducing agent, such as sodium(triacetoxy) borohydride was then added and stirring continued undernitrogen until the reaction was complete (as judged by hplc or tlc). Theresulting mixture was washed with an aqueous basic solution (e.g. sodiumor potassium carbonate) and extracted with a suitable solvent, e.g.dichloromethane. The dried organic phase was evaporated and the residuepurified either by column chromatography or by Bond Elut™ cartridge. Ifdesired, the isolated material was then converted into the hydrochloridesalt e.g. by treatment with ethereal hydrogen chloride.

Example 1

Preparation ofN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamineand di-hydrochloride and ditosylate salts thereof

(a) Preparation of 4-Hydroxy-7-iodoquinazoline

5-Iodoanthranilic acid (25 g) and formamide (200 mL) was combined andwarmed to 190° C. for 2 hours. After 15 minutes the reaction appeared tocompletely dissolve. The reaction was cooled down to room temperature,poured into water (500 mL) and allowed to stand for 1–2 hours. Thedesired product was collected by filtration.

(b) Preparation of 4-Chloro-7-iodoquinazoline

7-Iodoquinazolin-4-one (0.46 g) was treated with phosphorous oxychloride(5 ml) at reflux under nitrogen for 2 hours. The mixture was cooled,evaporated and partitioned between saturated aqueous sodium carbonateand ethyl acetate. The organic phase was dried and concentrated in vacuoto give the title compound (0.43 g); m/z (M+1+) 291.

(c) 4-(3-fluorobenzyloxy)-3-chloro nitrobenzene

2-chloro-4-nitrophenol (9.02 g, 52 mmol)), 3-fluorobenzyl bromide (9.85g, 52 mmol) and acetonitrile (90 mL) was combined at room temperatureunder nitrogen. Potassium carbonate (7.9 g, 57 mmol)) was added and thereaction mixture stirred at 60° C. for 2 hours and then cooled to roomtemperature. After cooling the reaction mixture was poured into water.The solids were collected by filtration and washed with diethyl ether toafford the desired product (13.98 g, 95% yield). [TLC system=1:1EtOAc:Hexanes, R_(f)=0.76]

(d) Preparation of 4-(3-fluorobenzyloxy)-3-chloro aniline

Under nitrogen, a Parr Shaker flask was charged with Pt/C 5% (135 mg)and ethanol (180 mL) and 4-(3-fluorobenzyloxy)-3-chloro nitrobenzene(13.5 g) was added. The reaction vessel was placed on a Parr ShakerApparatus under 25 psi of H₂ for 50 minutes. The catalyst was thenremoved by filtration through Celite and the filtrate concentrated toafford a light grey solid. The filtrate was triturated with diethylether and the solids collected by filtration (12.05 g, ˜100% yield).

(e) 6-Iodo-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-quinazolin-4-yl)amine

4-(3-fluorobenzyloxy)-3-chlorophenyl)-amine (12.3 g, 49 mmol),4-chloro-6-iodoquinazoline 14.2 g, 49 mmol) and isopropanol (250 mL)were combined and the reaction mixture was heated to 70° C. for 3.5hours. The resultant bright yellow solid product was collected byfiltration (25.5 g, 96/% yield). ¹H NMR (DMSO-d6) δ 9.83 (s, 1H); 8.92(s, 1H); 8.58 (s, 1H); 8.09 (d, 1H); 8.00 (d, 1H); 7.61 (d, 1H); 7.52(d, 1H); 7.44 (m, 1H); 7.20–7.33 (m, 3H); 7.15 (m, 1H); 5.21 (s, 2H); MSm/z 506 (M+1)

(f) Preparation of5-(4-{3-chloro-4-(3-fluorobenzyloxy)-anilino}-6-quinazolinyl)-furan-2-carbaldehyde

The title compound was prepared according to Procedure B followed byProcedure C from6-Iodo-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-quinazolin-4-yl)amine (1.0g, 1.82 mmol) and (1,3 dioxolan-2-yl)-2-(tributylstannyl)furan (11.17 g,2.73 mmol). ¹H NMR 400 MHz (DMSO-d6) δ12.05 (s, 1H); 9.68 (s, 1H); 9.43(s, 1H); 8.95 (s, 1H); 8.53 (d, 1H); 7.99 (D, 1H); 7.92 (s, 1H); 7.78(m, 1H); 7.66 (m, 1H); 7.63 (m, 1H); 7.47 (m, 1H); 7.40–7.30 (M, 3H);7.19 (m, 1H); 5.31 (s, 2H); MS m/z 472 (M+H).

(g) Preparation ofN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine

The title compound was prepared according to Procedure D as follows. The5-(4-{3-chloro-4-(3-fluorobenzyloxy)-anilino}-6-quinazolinyl)-furan-2-carbaldehyde(12.58 g, 24.7 mmol) was added to a solution of2-methanesulphonyl-ethylamine (4.55 g, 37.0 mmol) and triethylamine(2.0ml, 27.2 mmol) in 125 ml of tetrahydrofuran and 125 ml of methanol.After stirring 3 hours, the solution was cooled in an ice bath andsodium borohydride(2.42 g, 64.0 mmol) was added in five portions over a20 minute period. The reaction mixture was stirred overnight at roomtemperature and then quenched with the dropwise addition of 250 ml ofwater. The solution was concentrated in vacuo to remove the organicsolvents and the residual oily solution was extracted with 1 l of ethylacetate. The organic solution was washed with 1M sodium hydroxide andbrine and then dried with sodium sulfate. The solution was concentratedin vacuo to a very small volume and solid crystallized out. Thesuspension was filtered with a small volume of ethyl acetate, washedwith ether and dried in a vacuum oven at 65 C to give 10.0 g(70%) offree base as an off white solid. ¹H NMR 400 MHz (DMSO-d6) δ 9.60 (bs,1H); 9.32 (bs, 1H); 8.82 (bs, 1H); 8.34 (d, 1H); 8.0 (s, 1H); 7.88 (d,1H); 7.74 (d 1H); 7.45 (m, 1H); 7.34-7.23 (m, 4H); 7.17 (m, 1H); 6.83(d, 1H); 5.27 (s, 2H); 4.42 (s, 2H); 3.59 (m, 2H); 3.40 (m, 2H, obscuredby waterpeak); 3.12 (s, 3H); MS m/581 (M+H⁺).

(h) Preparation of5-(4-[3-chloro-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehydetosylate

A 2 liter, 3 neck round bottom flask equipped with a mechanical stirrerwas charged with 74.95 grams of the HCl salt of5-(4-[3-chloro-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehyde(prepared according to the Procedure D and Examples 1 (a)–(g), and 749.5mL THF. To this slurry was charged 84.45 mL of 2M NaOH and the reactantswere stirred for 30 minutes. The layers were separated and then theorganic layer was washed with 160 mL of H₂O. The organic layer wasslurried with 3.75 grams of Darco G60 and filtered through celite. Thefiltrate was collected and slowly added to 33.54 grams oftoluenesulfonic acid monohydrate with rapid stirring. The solids slowlyprecipitated out at ambient temperature. The mixture was cooled to 0° C.and stirred for 10 minutes. The mixture was filtered and pulled dry witha rubber dam, then dried in vacuo at 50° C. overnight. The yield of5-(4-[3-chloro-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehydetosylate was 84.25 grams (88.8%).

(i) Preparation ofN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamineditosylate anhydrate

To a 20 L reactor was added 13.3 vol of THF followed by 0.62 wt (2.93mol) of NaBH(OAc)₃. The 20 L reactor was set to maintain contents at 20°C. A second 20 L reactor was charged with 1000 grams, (1.55 mol) of5-(4-[3-chloro-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehydetosylate prepared by the procedure of Example 1 and 6.7 vol of THF. Tothe THF solution of5-(4-[3-chloro-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehydetosylate was added 0.325 vol (1.86 mol) diisopropylethylamine followedby 0.32 wt of 2-(methylsulfone)ethylamine, (321 g, 2.6 mol) and 0.15 volof IPA. After 1 hour, the preformed imine/THF solution was transferredby vacuum to the stirred suspension of NaBH(OAc)₃ in the first 20 Lreactor over 10 minutes. After 90 minutes, 4 vol of 5N NaOH was addedover 40 minutes via a pump. This solution was allowed to stir for 15minutes after which the stirrer was switched off and the layers wereallowed to separate. The aqueous layer was drained from the bottom ofthe reactor and the organic layer transferred to the empty 20 L reactorthrough a teflon-lined stainless steel jacketed transfer hose outfittedwith an in-line 0.45 μm filter. To this solution was added a 2 vol THFsolution of 4 wt (1180 g, 6.2 mole) of p-toluenesulfonic acidmonohydrate over 5 minutes. A yellowish precipitate was observed to comeout of solution and this was allowed to stir at room temperature for 12hours. The reaction was drained from the bottom of the reactor andfiltered through a ceramic filter lined with paper. The yellow filtercake was washed with 1 vol of a 95:5 THF/water solution and allowed toair dry overnight. After suctioning dry for 12 hours, the yellow filtercake was transferred to two glass trays and placed in the drying oven(42° C.) under house vacuum (18 in Hg) with a nitrogen bleed. The twoglass trays were removed from the oven and allowed to cool to roomtemperature and sampled accordingly. The isolated yield ofN-{3-Chloro-4[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methane-sulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamineditosylate (anhydrate) was 1264 grams (1.3 wt, 88%; 1443 g Th) and was ayellow solid.

Approximately 50 mg of the product was transferred to a Karl FisherVolumetric Moisture Apparatus (model DL35, Mettler, Hightstown, N.J.),which was operated according to the manufacturer's instructions. Theanhydrate water content was determined to be 0.31%.

(j) Preparation ofN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamineditosylate monohydrate (monohydrate form of compound of formula II)

A 20 L reactor was charged with 1 wt (930 g, 1.0 mol) ofN-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamineditosylate anhydrate prepared using the procedure of Example 2. To thiswas added 10 volumes of a pre-mixed 8:2 THF:deionized water solution andthe reactor was heated to 65° C. Complete dissolution was observed at50° C. The clear reaction mixture was transferred to another 20 Lreactor through a stainless steel jacketed transfer hose that wasequipped with an in-line 5.0 μm cartridge filter. The empty 20 L reactorand the filter line were washed with 0.2 vol of the pre-mixed 8:2THF:deionized water solution. An additional 1 vol of pre-mixed 8:2THF:deionized water solution was used to wash the material into thereaction mixture. The 20 L reactor was heated to ˜80° C. The reactiontemperature was then ramped down to 55° C. over 2 hours and then to 45°C. over 10 hours. After 10 hours, the temperature was adjusted to 25° C.and the reaction mixture allowed to stir at room temperature for 45minutes. The yellow precipitate was drained from the bottom of the 20 Lreactor into a ceramic filter lined with paper. The flow was fast andsmooth and the filter rate very good. The yellow filter cake was washedwith 0.6 volumes of a pre-mixed 8:2 THF:deionized water solution and theyellow solid was air dried for 4 hours and placed into a glass tray. Theglass tray was placed in a vacuum oven under house vacuum (˜18 in Hg) at60° C. with a nitrogen bleed for 2 days. After removal from the oven,the material was sampled accordingly. The yield was 743 grams (0.8 wt,80% o; 930 g th) as a bright yellow, crystalline solid.

Approximately 50 mg of the product was transferred to a Karl FisherVolumetric Moisture Apparatus (model DL35, Mettler, Hightstown, N.J.),which was operated according to the manufacturer's instructions. Themonohydrate water content was determined to be 1.99%, which is inagreement with the theoretical value of 1.92%.

Example 2

Preparation of(4-(3-Fluoro-benzyloxy-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)-quinazolin-4-yl)-amineand di-hydrochloride and ditosylate salts thereof

(a) Preparation of 2-Bromo-4-nitrophenol

2-Bromo-4-nitroanisole (20 g, 0.086 mol) was dissolved in DMF (414 mL)at room temperature under N₂. Sodium ethylthiolate (17.4 g, 0.207 mol)was added and the reaction mixture was warmed to 115° C. for 2 hours.The reaction was cooled to room temperature and diluted with EtOAc (200mL) and 1 M HCl (aq., 200 mL). The phases were separated, and thedesired product was extracted into 1 M NaOH (aq, 150 mL×3). The basicaqueous extracts were combined and acidified using conc. HCl. Thedesired product was extracted from the acidic aqueous solution usingEtOAc (250 mL×2). The combined organic layers were washed with brine anddried over sodium sulfate. The volatiles were removed in vacuo to afforda light brown semi-solid (9.8 g, 52% yield). ¹H NMR (DMSO-d6) δ 8.33 (m,1H); 8.09 (m, 1H); 7.07 (d, 1H).

(b) Preparation of 2-Bromo-1-(3-fluorobenzyloxy)-4-nitrobenzene

2-Bromo-4-nitrophenol (4.86 g, 0.0223 mol), triphenylphosphine (7.6 g,0.0290 mol), 3-fluorobenzylalcohol (3.65 g, 0.0290 mol) were combinedand dissolved in THF (89 mL). The reaction temperature was cooled to 0°C. and DIAD (4.50 g. 0.0290 mol) was added. The reaction was allowed towarm slowly to room temperature and stirred for 3 hours before it wasdiluted with water (1.00 mL) and EtOAc (100 mL). The layers wereseparated and the aqueous layer was extracted with EtOAc (200 mL×2). Theorganic extracts were combined and washed with brine, followed by dryingover sodium sulfate. The volatiles were removed in vacuo and theresidual semi-solid was treated with diethyl ether. The solids wereremoved by filtration. The volatiles from the resulting filtrate wereremoved in vacuo and the material was purified using EtOAc:Hexanes(90/10) in a biotage LC system to afford the title compound as a yellowsolid (3.73 g. 68% yield). ¹H NMR (DMSO-d6) δ 8.43 (d, 1H); 8.26 (m,1H); 7.45 (m, 1H); 7.38 (d, 1H); 7.30 (m, 2H); 7.17 (m, 1H); 5.39 (s,2H).

(c) Preparation of 3-Bromo-4-(3-fluorobenzyloxy)-aniline

Under a blanket of N₂, Pt/C (5%, 0.37 g) was charged to a Parr ShakerFlask. Ethanol (150 mL) and 2-bromo-1-(3-fluorobenzyloxy)-4-nitrobenzene(3.73 g, 0.011 mol) were added and the reaction mixture was placed on aParr Shaker Apparatus under 30 psi of H₂ for 5 h. The reaction wasfiltered through a pad of elite to remove the catalyst and the volatileswere removed from the filtrate. The residue was dissolved in the CH₂Cl₂(5 mL) and treated with conc. HCl (1 mL). The precipitate was collectedby filtration and free-based using saturated aqueous sodium bicarbonate(2.27 g, 67% yield) ¹H NMR (DMSO-d6) δ 7.4 (m, 1H); 7.23 (m, 2H); 7.11(m, 1H); 6.86 (d, 1H); 6.77 (m, 1H); 6.48 (m, 1H); 5.0 (s, 2H); 4.93(bs, 2H).

(d) Preparation of6-Iodo-(4-(3-fluorobenzyloxy)-3-bromophenyl)-quinazolin-4-yl)amine

The title compound was prepared according to Procedure A from3-bromo-4-(3-fluorobenzyloxy)-aniline (0.79 g, 2.7 mmol) and4-chloro-6-iodo-quinazoline (0.8 g, 2.7 mmol). ¹H NMR (DMSO-d6) δ 11.1(bs, 1H); 9.10 (s, 1H); 8.87 (s, 1H); 8.29 (d, 1H); 8.03 (s, 1H); 7.68(m, 1H); 7.62 (d, 1H); 7.45 (m, 1H); 7.33–7.26 (m, 3H); 7.16 (m, 1H);5.28 (s, 2H).

(e) Preparation5-(4-(3-Bromo-4-(3-fluorobenzyloxy)-anilino)-quinazolin-6-yl)-furan-2-carbaldehyde

The title compound was prepared according to Procedure B followed byProcedure C from6-iodo-(4-(3-fluorobenzyloxy)-3-bromophenyl)-quinazolin-4-yl)amine (1.0g, 1.82 mmol) and (1,3 dioxolan-2-yl)-2-(tributylstannyl)furan (1.17 g,2.73 mmol). ¹H NMR (DMSO-d6) δ 11.89 (bs, 1H); 9.66 (s, 1H); 9.41 (s,1H); 8.90 (s, 1H); 8.49 (d, 1H); 8.05 (m, 1H); 7.96 (d, 1H); 7.75 (m,1H); 7.70 (m, 1H); 7.61 (m, 1H); 7.43 (m, 1H); 7.30 (m, 3H); 7.16 (m,1H); 5.29 (s, 2H).

(f) Preparation of(4-(3-Fluoro-benzyloxy-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)-quinazolin-4-yl)-aminedihydrochloride

The title compound was prepared according to Procedure D from a mixtureof5-(4-(3-Bromo-4-(3-fluorobenzyloxy)-anilino)-quinazolin-6-yl)-furan-2-carbaldehyde(0.623 g, 1.2 mmol) in dichloroethane (12 mL),triethylamine (0.167 mL,1.2 mmol), acetic acid (0.216 mL 3.6 mmol), and2-methanesulphonylethylamine (0.447 g, 3.6 mmol). The reaction mixturewas warmed to reflux for 1 hour and then cooled to rt before addingsodium triacetoxyborohydride (0.5 g). After 0.5 hours of stirring,another aliquot of sodium triacetoxyborohydride (0.5 g) was added andthe reaction was stirred an additional 0.5 hours. The reaction wasquenched by the addition of a saturated solution of sodium bicarbonate(aq, 50 mL). EtOAc (50 mL) was added and the layers were separated. Theorganics were washed with brine and dried over sodium sulfate. Thevolatiles were removed in vacuo. Purification of the compound wasachieved using Biotage column chromatography; eluents: CH2Cl2, EtOH,Et3N (150:8:1). The appropriate fractions were combined and thevolatiles were removed in vacuo. The compound was crystallized fromEtOAc and Et₂O to afford a yellow solid. The hydrochloride salt was madeby dissolving the material in a minimal amount of EtOAc and adding 2MHCl in diethyl ether (0.5 mL) to afford a dark yellow solid (0.27 g, 35%yield). ¹H NMR (DMSO-d6) δ 11.70 (bs, 1H); 9.84 (bs, 2H); 9.59 (s, 1H);8.89 (s, 1H); 8.39 (d, 1H); 8.14 (s, 1H); 7.93 (d, 1H); 7.80 (d, 1H);7.45 (m, 1H); 7.31 (m, 4H); 7.16 (m, 1H); 6.83 (m, 1H); 5.30 (s, 2H);4.43 (s, 2H); 3.67 (m, 2H); 3.40 (m, 2H); 3.12 (s, 3H).

(g) Preparation of(4(3-Fluoro-benzyloxy)-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)quinazolin-4-yl)-amineditosylate.

The HCl salt of5-(4-[3-bromo-4-(3-fluorobenzyloxy)-anilino]-6-quinazolinyl)-furan-2-carbaldehyde,is prepared according to Procedure D and Example 1(e), and is convertedto the tosylate salt according to the procedure of Example 1(h). Theresultant carbaldehyde tosylate product is used to prepare the(4-(3-Fluoro-benzyloxy)-3-bromophenyl)-(6-(5-((2-methanesulphonyl-ethylamino)-methyl)-furan-2-yl)quinazolin-4-yl)-amineditosylate according to the procedure of Example 1(i).

Example 3

Preparation of(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-amineand di-hydrochloride and ditosylate salts thereof

(a) Preparation ofN-(4-(3-fluorobenzyloxy)-chlorophenyl)-6-(1-ethoxyvinylether)-quinazolin-4-yl)-amine

To a suspension of the6-iodo-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-quinazolin-4-yl amine(12.6 g, 24.93 mmol) in acetonitrile (100 mL) was addedtributyl(1-ethoxyvinyl)stannane (9 g, 24.93 mmol) andbis(triphenylphosphine) palladium (II) chloride (1.75 g, 2.29 mmol). Thereaction mixture was refluxed for 18 hours, then filtered through a plugof silica gel. The resulting solution was poured into 5% aqueous NH₄OH(200 mL) and extracted with ethyl acetate (500 mL). The organic layerwas dried over anhydrous sodium sulfate, concentrated, and purified bysilica gel chromatography to provide the title compound as a yellowsolid (7.2 g, 64% yield). ¹H NMR (400 MHz, d₆ DMSO) δ 9.92 (s, 1H), 8.76(s, 1H), 8.58 (s, 1H), 8.08 (m, 1H), 8.01 (m, 1H), 7.76 (m, 2H), 7.48(m, 1H), 7.32 (m, 3H), 7.22 (m, 1H), 5.28 (s, 2H), 5.02 (s, 1H), 4.56(s, 1H), 4.01 (q, 2H), 1.42 (t, 3H); ESI-MS m/z 449.9 (M+H)⁺.

(b) Preparation ofN-{4-[(3-fluorobenzyloxy)]-chlorophenyl}-6-[2-({[2-(methanesulphonyl)ethyl]-[trifluoroacetyl]amino}methyl)-1,3-thiazol-4-yl]-quinazolin-4-yl)-amine

To a solution ofN-(4-(3-fluorobenzyloxy)-chlorophenyl)-6-(1-ethoxyvinylether)-quinazolin-4-yl)-amine(7.1 g, 15.8 mmol) in a THF (150 mL)/H₂O (5 mL) mixture cooled to 0° C.was added N-bromosuccinimide (2.81 g, 15.8 mmol). The resulting mixturewas stirred for 0.25 hours, then dried over anhydrous sodium sulfate andconcentrated. The crudeN-(4-(3-fluorobenzyloxy)-chlorophenyl)-6-(bromomethylketone)-quinazolin-4-yl)-amineand N-(trifluoroacetyl)-N-(methanesulphonylethyl)-aminomethylthioamide(4.61 g, 15.8 mmol) were dissolved in DMF (50 mL) and heated at 70° C.for 1 hour. The reaction mixture was concentrated, then diluted withdichloromethane (300 mL) and washed with saturated sodium bicarbonatesolution (100 mL). The organic layer was dried over anhydrous sodiumsulfate, concentrated, and purified by silica gel chromatography toprovide the title compound as a foam (4.6 g, 42% yield). ESI-MS m/z694.1 (M+H)⁺.

(c) Preparation ofN-{4-[(3-fluorobenzyloxy)]-chlorophenyl}-6-[2-({[2-(methanesulphonyl)ethyl]-amino}methyl)-1,3-thiazol-4-yl]-quinazolin-4-yl)-aminehydrochloride

To a solution ofN-{4-[(3-fluorobenzyloxy)]-chlorophenyl}-6-[2-({[2-(methanesulphonyl)ethyl]-[trifluoroacetyl]amino}methyl)-1,3-thiazol-4-yl]-quinazolin-4-yl)-amine(4.6 g, 6.63 mmol) in methanol (100 mL) was added 2M NaOH (50 mL). Theresulting mixture was stirred at room temperature for 2 hours,concentrated to ½ volume, poured into H₂O (100 mL), and extracted withdichloromethane (300 mL). The organic layer was dried over anhydroussodium sulfate, concentrated, and purified by silica gel chromatography.The resulting amine was dissolved in dichloromethane/methanol (3:1, 100mL) and then 4M HCl/dioxane (20 mL) was added. The resulting mixture wasconcentrated and filtered to provide the title compound as a yellowsolid (4.0 g, 90% yield). ¹H NMR (400 MHz, d₄ MeOH) δ 9.38 (s, 1H), 8.82(s, 1H), 8.78 (d, 1H), 8.36 (s, 1H), 7.94 (s, 1H), 7.92 (d, 1H), 7.63(m, 1H), 7.41 (m, 1H), 7.26 (m, 1H), 7.22 (m, 2H), 7.04 (m, 1H), 5.24(s, 2H), 4.82 (s, 2H), 3.84 (m, 2H), 3.76 (m, 2H), 3.12 (s, 3H); ESI-MSm/z 597.1 (M+H)⁺.

(d) Preparation of(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-amineditosylate.

The HCL salt of(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-aminewas prepared according to Procedures 3(a) to (c) and then converted tothe(4-(3-Fluoro-benzyloxy)-3-chlorophenyl)-(6-(2-((2-methanesulphonyl-ethylamino)-methyl)-thiazol-4-yl)quinazolin-4-yl)-amineditosylate salt according to the procedure of Examples 1 and 2. ¹H NMR(300 MHz, d6-DMSO) 11.4 (br s, 1H), 9.51 (br s, 1H), 9.24 (s, 1H), 8.95(s, 1H), 8.68 (d, J=9 Hz, 1H), 8.42 (s, 1H), 7.96 (d, J=9 Hz, 1H), 7.89(d, J=2 Hz, 1H), 7.64 (dd, J=2, 9 Hz, 1H), 7.47 (m, 5H), 7.34 (m, 3H),7.20 (t, J=9 Hz, 1H), 7.10 (d, J=8 Hz, 4H), 5.32 (s, 2H), 4.76 (d, 2H),3.61 (s, 4H), 3.15 (s, 3H), 2.28 (s, 6H).

Biological Data

Tumor Studies: HN5

HN5 cells were cultured in RPMI 1640+10% Fetal bovine serum, Sodiumpyruvate and L-Glutamine at 370° in a 95/5% air/CO₂ atmosphere. Cellswere harvested following trypsin digestion and brought to a density of2×10⁶ cells/200 μl in PBS. Tumors were initiated by injection of thecell suspension subcutaneously in the axillary region.

Tumor Studies: BT474

The BT474 xenograft was maintained by serial transplantation in SCIDmice. Tumors are initiated by injection of tumor fragmentssubcutaneously in the axillary region.

Tumor Studies: NCl H322

NCl H322 cells were cultured in RPMI 1640+10% Fetal bovine serum, Sodiumpyruvate and L-Glutamine at 37° in a 95/5% air/CO₂ atmosphere. Cellswere harvested following trypsin digestion and brought to a density of2×10⁶ cells/200 μl in PBS. Tumors were initiated by injection of thecell suspension subcutaneously in the axillary region. In addition someexperiments were performed following serial transplantation of tumorfragments in SCID mice. Tumors were initiated by injection of tumorfragments subcutaneously in the axillary region.

Tumor Studies: Measurements

For the xenograft models used here solid tumors were measured byelectronic caliper measurement through the skin, measurements weretypically made twice weekly. In the examples presented, tumors weremonitored beyond the duration of therapy

Tumor Studies: Formulation and Administration

Drugs were administered by P.O. or I.V. routes. The title compound orsalt thereof of Example 1 was formulated in aqueous 0.5% hydroxypropylmethylcellulose, 0.1% Tween 80 and administered as a suspension twicedaily for 21 days as indicated in the respective figures. Taxol®(Bristol Myers Squibb Co.) was purchased preformulated in Cremophor-ELsaline and diluted into saline to a final Cremophor-EL concentration of5 or 10% Cremophor-EL for 10 or 20 mg/kg Taxol therapy respectively.Taxol was administered I.V., once a day, for 5 days (days 1–5 of Example1 therapy) as indicated in the respective figures. Carboplatin (Sigma)was formulated in saline and was administered I.V., once a day, for two5 day periods as indicated in the respective figure (days 1–5 and 15–19of Example 1 therapy). These studies were performed under IACUC # 468.The results are illustrated in FIGS. 1–3.

FIG. 1 illustrates dosing of a HN5 (head and neck) subcutaneous humanxenograft mouse model with the compound of Example 1 and/or carboplatin.Carboplatin as a montherapy in the HN5 s.c. human xenograft mouse modelshowed some anti-tumor activity (about 45% tumor growth inhibition athighest dose). Dosing of the compound of Example 1 also showedanti-tumor activity in the same model as monotherapy (about 80% tumorgrowth inhibition at highest dose). When the compound of Example 1 andcarboplatin were used in combination, 100–120% tumor growth inhibitionwas observed during treatment

FIG. 2 illustrates dosing of a BT474 (breast) sub cutaneous humanxenograft mouse model with the compound of Example 1 and/or Taxol®.Taxol® dosed as monotherapy in the BT474 s.c. human xenograft mousemodel showed some anti-tumor activity (about 45% tumor growthinhibition). The compound of Example 1 also showed anti-tumor activityin the same model when dosed as monotherapy (about 90% tumor growthinhibition at the highest dose tested). When the compound of Example 1and Taxol® were used in combination, 100–120% anti-tumor activity wasobserved during treatment. There was a significant delay in tumorre-growth observed, tumor growth inhibition was sustained forapproximately 40 days after treatment ceased.

FIG. 3 illustrates dosing of a NCl H322 (lung) sub cutaneous humanxenograft mouse model with the compound of Example 1 and/or Taxol®.Taxol® dosed as monotherapy in the NCl H322 s.c. human xenograft mousemodel showed some anti-tumor activity (about 45% tumor growthinhibition). The compound of Example 1 also showed anti-tumor activityin the same model when dosed as monotherapy (about 100–130% tumor growthinhibition). When the compound of Example 1 and Taxol® were used incombination, tumor growth was completely arrested (below detection—notmeasurable). There was a significant delay in tumor re-growth observedafter treatment ceased.

1. A method of treating breast cancer in a mammal, comprising:administering to said mammal a therapeutically effective amount of: (a)a compound of formula II:

 salts, or solvates; and (b) paclitaxel.
 2. The method of claim 1,wherein said method comprises administering to said mammal atherapeutically effective amount of (a) a compound of formula II,

 or salt thereof; and (b) paclitaxel.